PDIA3 / ERp57 Recombinant Rabbit Monoclonal Antibody [PSH08-79] - BSA and Azide free
Catalog# HA751244
PDIA3 / ERp57 Recombinant Rabbit Monoclonal Antibody [PSH08-79] - BSA and Azide free
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WB
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IHC-P
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FC
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IF-Cell
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Human
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Mouse
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Rat
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HA723031
含抗保成分
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unconjugated
Overview
Product Name
PDIA3 / ERp57 Recombinant Rabbit Monoclonal Antibody [PSH08-79] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human PDIA3 aa 120-370
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, FC, IF-Cell
Molecular Weight
Predicted band size: 57 kDa
Positive Control
HepG2 cell lysate, HeLa cell lysate, HEK-293 cell lysate, U-87 MG cell lysate, RAW264.7 cell lysate, mouse liver tissue lysate, rat liver tissue lysate, U-87 MG, NIH/3T3, C6, human liver tissue, human placenta tissue, mouse liver tissue, mouse placenta tissue.
Conjugation
unconjugated
Clone Number
PSH08-79
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IHC-P
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1:200-1:1,000
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FC
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1:1,000
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IF-Cell
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1:100
Target
Function
The PDIA3 protein is a thiol oxidoreductase that has protein disulfide isomerase activity. PDIA3 is also part of the major histocompatibility complex (MHC) class I peptide loading complex, which is essential for formation of the final antigen conformation and export from the endoplasmic reticulum to the cell surface. This protein of the endoplasmic reticulum interacts with lectin chaperones such as calreticulin and CNX in order to modulate the folding of proteins that are newly synthesized. It is believed that PDIA3 plays a role in protein folding by promoting the formation of disulfide bonds, and that CNX facilitates the positioning substrates next to the catalytic cysteines. This function allows it to serve as a redox sensor by activating mTORC1, which then mediates mTOR complex assembly to adapt cells to oxidative damage. Thus, PDIA3 regulates cell growth and death according to oxygen concentrations, such as in the hypoxic microenvironment of bones. Additionally, PDIA3 activates cell anchorage in bones by associating with cell division and cytoskeleton proteins, such as beta-actin and vimentin, to form a complex which controls TUBB3 folding and proper attachment of the microtubules to the kinetochore. PDIA3 also plays a role in cytokine-dependent signal transduction, including STAT3 signaling.PDIA3 may also participate in Vitamin D (specifically, calcitriol) signaling as a membrane-bound receptor.
Background References
1. [1]Basu A ,Ross C K C ,Rios-Colon L , et al.LEDGF/p75 Overexpression Attenuates Oxidative Stress-Induced Necrosis and Upregulates the Oxidoreductase ERP57/PDIA3/GRP58 in Prostate Cancer.[J].PLoS ONE,2017,11(1):e0146549.
2. [2]Luo H J ,Wang X F ,Zhao W J , et al.PDIA3 defines a novel subset of adipose macrophages to exacerbate the development of obesity and metabolic disorders.[J].Cell metabolism,2024,36(10):2262-2280.e5.
Subcellular Location
Endoplasmic reticulum Endoplasmic reticulum lumen Melanosome
Synonyms
58 kDa glucose regulated protein antibody
58 kDa glucose-regulated protein antibody
58 kDa microsomal protein antibody
Disulfide isomerase ER 60 antibody
Disulfide isomerase ER-60 antibody
Endoplasmic reticulum resident protein 57 antibody
Endoplasmic reticulum resident protein 60 antibody
ER p57 antibody
ER protein 57 antibody
ER protein 60 antibody
ExpandImages
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Western blot analysis of PDIA3 / ERp57 on different lysates with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/2,000 dilution.
Lane 1: HepG2 cell lysate
Lane 2: HeLa cell lysate
Lane 3:HEK-293 cell lysate
Lane 4: U-87 MG cell lysate
Lane 5:RAW264.7 cell lysate
Lane 6:mouse liver tissue lysate
Lane 7: rat liver tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 57 kDa
Observed band size: 57 kDa
Exposure time: 6 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751244) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of PDIA3 / ERp57 on different lysates with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/1,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-PDIA3 / ERp57 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 57 kDa
Observed band size: 57 kDa
Exposure time: 6 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751244) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of U-87 MG cells labeling PDIA3 / ERp57 with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of NIH/3T3 cells labeling PDIA3 / ERp57 with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of C6 cells labeling PDIA3 / ERp57 with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751244) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751244) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751244) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse placenta tissue with Rabbit anti-PDIA3 / ERp57 antibody (HA751244) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751244) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of U-87 MG cells labeling PDIA3 / ERp57.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA751244, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of NIH/3T3 cells labeling PDIA3 / ERp57.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA751244, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of C6 cells labeling PDIA3 / ERp57.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA751244, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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